Objective To discuss the influence of intravenous anesthesia with procaine on the release of central nervous Ach. Methods 50 samples of plasma were collected from healthy people, and 30 samples of plasma from cases prior to surgical operation, during anesthesia and after operation respectively. The activity of Ach in the plasma was measured. Then observe the dropping speed and concentration of procaine at different time during anesthesia, and the subsequent changes of the activity of plasma Ach and the slowdown of its activity. Results The data showed that the activity of plasma Ach prior to operation was considerably lower than that of the control group (P

Objective To construct the stably over-expressing hB7-H3 gene human pancreatic cancer cell line PANC1,and provide tools for further investigating the function of hB7-H3.Methods hB7-H3 gene fragment was inserted into lentiviral plasmid GV287 carrying GFP to construct recombinant hB7-H3-GV287 plasmid vector.293T cells were transfected,and the GFP expression was evaluated under fluorescence microscopy.Western Blot wasused to detect the expression of hB7-H3 protein.Lentiviral vectors were packaged and the titer was determined.The recombinant hB7-H3 expressed lentivirus was used to infect PANC1 cell.Flow cytometry was applied to detecte the percentage of GFP and hB7-H3 positive cells.Real-time PCR and Western Blot was used to verify the mRNA and protein expression of hB7-H3.Self-cyclizing GV287 plasmid served as negative control (NC).Results PCR amplified fragment of recombinant plasmid was around 1 368 bp,and no amplified production of NC plasmid was observed.The DNA sequencing of recombinant plasmid was completely consistent with the designed fragment,indicating that hB7 H3-GV287 plasmid was successfully constructed.293T cells transfected with recombinant plasmid expressed hB7-H3 protein,while those cells transfected with NC plasmid did not express hB7-H3 protein.The virus titer of lentiviral packaged recombinant hB7-H3 plasmid was 2 × 108 TU/ml.The percentage of hB7-H3 positive cells,hB7-H3 mRNA and protein expression in PANC1 cells infected with cells infected with hB7-H3 lentivirus was 94.3％,5.09 ± 0.24 and 2.85 ± 0.27,respectively,which was obviously higher than 18.5％,1.28 ± 0.53 and 0.44 ± 0.69 in cells infected with NC lentivirus,and the differences were statistically significant (P value ＜0.01).Conclusions A human pancreatic cancer cell line PANC1 stably over-expressing hB7-H3 was successfully constructed.

Objective To observe the inhibition effect of selective cyclooxygenase-2 inhibitor (celecoxib) on the experimental choroidal neovascularization (CNV).Methods Thirty 8-10 weeks old healthy male Brown-Norway(BN) rats were randomly divided into the control,laser and celecoxib group,with 10 rats in each group.At the dosage of 50 mg/kg,celecoxib was garaged twice per day.After 7 days,experimental CNV was induced by Krypon laser on laser group and celecoxib group.Fundus fluorescein angiography (FFA) was performed on days 3,7,14,21,30 after laser photocoagulation.On days 21 after photocoagulation,5 rats in each group were sacrificed and the relative thickness of CNV membranes,the expression of COX-2,vascular endothelial growth factor(VEGF) and matrix metalloproteinase-2 (MMP-2) Were studied by histopathologic or immunohistochemistry examination.Results On days 21 after photocoagulation,the incidence of CNV in the celecoxib group is significantly lower than that in the laser group (X~2=7.1068,P=0v0077);the relative thickness of the CNV membranes in the celecoxib group is reduced 41.38% compared to the laser group,the difference is statistically significant (t=16.7600,P=0.0000).COX-2,VEGF and MMP-2 expression in the CNV membrane of celecoxib group were significantly lower than in control group (t=5.7100,5.8400,8.0200;P=0.0000);the COX-2,VEGF and MMP-2 expressions in choroid and retina of control group were weak.Conclusion Prophylactic celecoxib can reduce the expression of VEGF and MMP-2 by inhibiting COX-2,and prevent the CNV induced by laser photocoagulation.

Objective To observe the clinical effects of different doses of Xuebijing injection in the treatment of sepsis. Methods Eighty patients with sepsis from First Center Clinical College of Tianjin Medical University were selected and randomly divided into four groups:the control group,Xuebijing small,middle and large dose groups(each, 20 patients). All patients in the four groups were given Western medicine conventional treatment,whereas in the Xuebijing small,middle and large dose groups,additionally they were intravenously dripped with Xuebijing injection 50 mL, 2 times a day,100 mL,2 times a day and 100 mL,4 times daily respectively. The patients' vital signs,blood routine, interleukin-18(IL-18),procalcitonin(PCT)and C- reactive protein(CRP)were detected before treatment(0 d)and 3,7 and 14 days after Xuebijing injection,28-day and 60-day death rate were observed. Results With prolonged treatment,the body temperature,heart rate(HR),respiratory rate(RR),white blood cell count(WBC),IL-18,PCT, CRP levels were all decreased in each group(all P<0.05),and on 14 days they reached the valley;when compared with middle,small dose Xuebijing groups and control group,in the Xuebijing large dose group they were decreased the most significantly over the same period〔temperature (℃):36.22±0.55 vs. 36.55±0.99, 36.87±1.05, 36.93±1.07,HR(beats/min):82.11±8.99 vs. 93.45±10.02,109.02±11.44,113.13±10.34,RR(breaths/min):15.48±6.45 vs. 17.33±6.94,20.35±7.19,22.34±6.67,WBC(×109/L):5.35±1.37 vs. 8.35±2.98,10.54±3.22, 13.33±4.78,IL-18(ng/L):107.36±55.89 vs. 136.55±80.19,177.78±102.32,193.93±98.07,PCT(μg/L):0.33±0.19 vs. 0.66±0.22,1.62±0.44,2.78±0.34,CRP(mg/L):15.40±1.37 vs. 48.34±8.98,100.45±30.67, 131.92±49.53〕,the differences all being statistically significant(all P<0.05). The 28-day and 60-day death rate in Xuebijing large dose group were significantly lower than those in control group,Xuebijing small,middle dose groups (28-day:25% vs. 40%,40%,45%;60-day:30% vs. 45%,40%,50%). Conclusion The time for onset of therapeutic effect in large dose Xuebijing group is short,and reduce the levels of inflammatory mediators significantly, and improve the prognosis of patients with sepsis,therefore it can be used clinically.

Objective: To observe the clinical efficacy of combined acupuncture and speech rehabilitation in treating postapoplectic aphasia. Methods: Sixty subjects were randomized into two groups. 30 cases in the treatment group were treated by both speech rehabilitation and acupuncture therapy, while 30 cases in the control group were treated by speech rehabilitation only. After 30-day treatments, the clinical efficacy was observed. Results: The total effective rate and speech score were higher in the treatment group than in the control group (P<0.05). Conclusion: Combined acupuncture and speech rehabilitation is quite effective for postapoplectic aphasia.

Objective To compare the performance of two commercially available ELISA kits for detection of diphtheria IgG antibody to provide data for the proper kit choice. Methods Thirty-three serum samples from staff of the capital airdrome and two standard sera were tested for the evaluation of the kits.Commercial ELISAs were compared with respect to their reproducibility(intra-assay variation and inter-assay variation)and accuracy.Results For the detection of standard serum,intra-assay variation(Coefficient of Variation,CV)was 13.5507% for IBL kit and 5.9541% for Virion kit,the inter-assay variation(CV)was 9.9890% for IBL and 2.4728% for Virion kit.For the serum sample,intra-assay variation(CV)was 14.5270% for IBL and 11.0612% for Virion,the inter-assay variation(CV)11.0039% for IBL and 8.1506% for Virion kit.For the detection of standard serum,inter-assay variation(CV)was 9.9890% for IBL and 2.4728% for Virion.The accuracy was analyzed by variation between the actual concentration and measured concentration of standard serum,the CV was 17.6889% for IBL and 1.3966% for Virion.Conclusion The intra-assay and inter-assay variation of the two kits are generally acceptable(CV

Past decade,plants as production systems for recombinant proteins has been actively investigated,and is one of the most active research fields in life sciences. Plants are attractive as protein factories because they with advantages of cost,scale,and safety,and emerge capability to produce pharmaceutical production with bioactivity proved by previously studies. The plant-made pharmaceuticals possess enormous viability for market and wide prospects for application. Based on those reasons,the rise of plant bioreactor was reviewed,and research advance on the expression systems and plant-made recombinant proteins was introduced. Bottleneck problems and adopted strategies using plant system to produce recombinant proteins and research hotspot in the near future were summarized,and prospect of plant bioreactor in future was described. Synchronously,some opinions including research progress of plant bioreactor,differences to foreign and strategies in the future were discussed.

Sleep plays an important role in the modulation of endocrine function and energy metabolism;hormone levels are markedly different during a sleepless night compared with a night of normal sleep.Sleep deprivation may change the levels of cortisol,growth hormone(GH)and thyrotropin(TSH)at night.In addition,sleep curtailment disturbs the balance of anorexi- genic(leptin)and orexigenic(ghrelin)factors,and does harmful to glucose tolerance.

Based on the high solubility of azodicarbonamide (ADC) in hot solution and its electrostatic interaction with Nafion film,a new electroanalytical method was developed for the determination of ADC by using Nafion film electrode.The effect of temperature on the solubility of ADC and the mechanism of the reduction reaction of ADC on Nafion film electrode were investigated.Under the experimental conditions such as water bath at a constant temperature of 80 ℃,pH 6.0 and optimal test parameters,the differential pulse voltammetric response was proportional to the concentration of ADC in the range of 0.93-10.5 μg/L,and the detection limit was estimated to 0.58 μg/L.The relative standard deviation was less than 5.86 ％ and the recovery was 95.8％-104.0％ for the determination of the ADC in flour samples.The semicarbazide and nitrofurazone did not interfere with the determination of ADC.

Objective To investigate the effect of has-miR-663 on proliferation and apoptosis of human adrenal cortical cell line SW-13.Methods Based on the first stage study,in adrenocorticotropinindependent macronodular adrenal hyperplasia (AIMAH),hsa-miR-663 was up-regulated.The synthetic hsa-miR-663 mimics and inhibitors were transfected the SW-13 cell line.Real-time quantitative PCR (RT-PCR) tested the effect of the transfection.According to the results of RT-PCR,cell line was divided into five groups,including blank group,mimics NC group,inhibitor NC group,mimics group,inhibitor group.Then proliferating of the cell was tested by MTT and the apoptosis by Annexin V-FITC/PI.Result By RT-PCR,mimics group showed that has-miR-663 was over-expressed,and inhibitor group were knocked down.MTTshowed that,compared with NC group and Blank group,SW-13 cell line proliferated faster in mimics group and slower in inhibitor group.Annexin V-FITC/PI showed that apoptosis of SW-13 cell line was not different in all the five groups.Conclusion Up-regulating has-miR-663 could accelerate proliferation of adrenal cortical cells.The microRNA might play some role in the pathogenesis of AIMAH.